This proposal requests funds for the purchase of a Zeiss LSM 510 confocal microscope system that will be dedicated to the cellular imaging needs of five NIH-funded Major Users located on the Bayview campus of the Johns Hopkins University. This campus contains the Johns Hopkins Bayview Medical Center Hospital, the Hopkins Geriatric Center and Asthma and Allergy Center, the renovated Baltimore City Hospital containing 60,000 square feet of renovated research laboratories, and three buildings housing the intramural research programs of the National Institute on Aging (NIA) and National Institutes in Drug Abuse (NIDA). The Hopkins Bayview faculty account for one-third of the total research dollars awarded to the Department of Medicine (~$50 million dollars in FY 2005), and collectively, has published more than 3,700 articles during the period of 2000-2005. There is a dire need for advanced optical imaging resources on this campus, yet there is no multi- user facility here. The closest such facility is 4.2 miles away at the Hopkins East Baltimore campus. Although a shuttle bus connects the two campuses, transferring samples downtown is cumbersome and impractical for live cell imaging, the focus of a number of investigators in this application. Although there are several confocal microscopes on campus that belong to the NIA and NIDA intramural research programs, there is no unencumbered mechanism for Hopkins Bayview investigators to access these instruments. The instrument will initially be dedicated to the needs of the five major users, whose research focuses on the molecular and cellular understanding of clinical immunology and allergic reactions as well as vascular, cardiac, pulmonary, and rheumatoid diseases. Requests for imaging from a minor user group that currently includes an additional 14 NIH-funded investigators on the Bayview campus will be accommodated as time on the instrument allows. The combined group has critical needs for (1) co-localization of proteins and/or lipids to each other and to sub- cellular organelles; (2) FRET-based methods for characterizing protein-protein interactions, intracellular kinase activity, pH, and calcium; (3) multi-fluorescent live cell imaging; and (4) multi-fluorescent 3-D imaging. The confocal imaging system requested not only meets these needs, it is readily capable of accommodating the future addition of spectral deconvolution hardware and software (META) as well as 2-photon capabilities. The instrument will serve as the cornerstone for an imaging facility to meet the current and future needs of investigators at Hopkins Bayview. RELEVANCE: Research of the combined user groups is focused on the molecular and cellular understanding of clinical immunology and allergic reactions, as well as vascular, cardiac, pulmonary, and rheumatoid diseases. Collectively, these diseases represent a significant burden on the health care system and account for a large fraction of deaths in the United States every year. [unreadable] [unreadable] [unreadable]